The main objectives of the Vector Core are to provide high quality self-complementary (sc) and single-stranded (ss) rAAV1 vectors and serologically screen NHP animals suitable for rAAV1 anti-viral studies. We will accomplish these goals through the following specific aims: Aim 1. To design, create, produce and quality control test scAAV1 vector lots at different scales with a variety of transgenes and expression cassettes to serve the specific needs of other investigators of this program project. More specifically, 120 rhesus macaques will be enrolled for different studies over 5 years and hundreds of mice will used for pre-macaque evaluation. In average, we estimate that 15-20 vector lots will be produced annually to meet the needs of those studies. Aim 2. To screen NHP populations for pre-existing immunity against AAV1 by using both in vitro and in vivo neutralizing antibody (NAB) assays to select AAV1-NAB free animals for vaccine and therapeutic studies. Pre-screening of NHP population to select the animals without preexisting neutralizing antibody (NAB) to rAAV1 is essential for rAAV1-mediated anti-HIV immunoadhesin gene transfer. Our data suggested that the serological prevalence of primate-derived AAVs in NHP populations ranges from 60-80%. This implies that we may have to screen more than 360 animals to identify 120 animals free of AAV1 NAB. Aim 3. To develop novel and scalable rAAV production method for larger scale translational NHP studies and future clinical development of rAAV1-based anti- HIV vaccine and therapeutics. Our current AAV production system should meet the vector needs in the early stage of this program project. However, large scale vector production may become a bottle neck for larger translational NHP studies and future clinical development as well. We will utilize our extensive experience in developing various vector packaging cell lines and infection-based vector production system to develop a 293 cell infection-based novel and scalable production method to overcome this limitation.